Inhibitory serum factor of lymphoproliferative response to allogeneic cells in pregnancy

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PAUL. ,0.

lfllD lC A L

S ilv ia D a h e r, F la v ia F o n s e c a , R o s e a n e M a tta r, _ C h lo e C a m b a M u s a tti, M a ria G e rb a s e d e L im a

I n h ib ito r y s e r u m f a c to r o f I y m p h o p r o lif e r a tiv e r e s p o n s e to

a llo g e n e ic c e lls in p r e g n a n c y

C l i n i c a i I m m u n o l o g y a n d R h e u m a t o l o g y D i s c i p l i n e o / t h e P e d i a t r i c s D e p a r t m e n t - U n i v e r s i d a d e F e d e r a l d e

S ã o P a u l o - E P M - S ã o P a u l o , B r a z i l

Introduction: An inhibitory serum factor of m ixed Iym phocyte culture (M LC) has been associated with successful pregnancy after.

Iym phocyte transfusion in wom en with unexplained recurrent spontaneous abortions (ASA). O bjective: Investigate whether the

inhibitory serum factor of M LC is essential for a successful pregnancy. M ethod: Sera from 33 healthy pregnant wom en and from 40 wom en with ASA were assessed by a one-way M LC in which the wom an's Iym phocytes were stim ulated with her partner's Iym phocytes or with third.party Iym phocytes. Results: An inhibitory serum effect (inhibition > 50% as com pared to norm al serum ) was detected in

45% of the pregnant wom en who had at least 1 previous parity, in8 0 /0 of the prim igravidea, in 29% of those with one abortion and in

58% of those with m ore than one abortion. Conclusion: ,M LC inhibitory serum factor does not seem to be an essential factor for

pregnancy developm ent. Therefore, it should not be considered as a param eter for the assessm ent of ASA patients.

UNITERM S: M ixed Iym phocyte culture teste Pregnancy. Habitual abortion.

INTRO DUCTIO N

T

h e m a t e r n a l i m m u n o l o g i c r e s p o n s e t o t h e f e t u s h a s b e e n a s s e s s e d i n s e v e r a l s t u d i e s .I-4 H o w e v e r ,

t h e e s s e n t i a l f a c t o r s t h a t l e a d t o t h e e s t a b l i s h m e n t

a n d m a i n t e n a n c e o f s u c c e s s f u l p r e g n a n c y r e m a i n u n c l e a r .

I t h a s b e e n s u g g e s t e d t h a t i n a n o r m a l p r e g n a n c y t h e r e i s a

m o d u l a t i o n o f p o t e n t i a l l y d e l e t e r i o u s i m m u n e r e s p o n s e s

a g a i n s t t h e f e t u s i n o r d e r t o p r e v e n t r e j e c t i o n r e a c t i o n s .5, 6

T h u s , a n a b n o r m a l a l l o g e n e i c r e s p o n s e , w i t h i n s u f f i c i e n t

p r o d u c t i o n o f b l o c k i n g f a c t o r s , h a s b e e n p r o p o s e d a s t h e

c a u s e o f c e r t a i n u n e x p l a i n e d r e c u r r e n t s p o n t a n e o u s

A d d re s s fo r c o rre s p o n d e n c e : S ilv ia D a h e r

R u a B e la C in tra , 1 9 2 0 a p t. 4 1

S ã o P a u lo / S P - B ra s il- C E P 0 1 4 1 5 -0 0 2

a b o r t i o n s ( R S A ) . 1 - 3 , 6I n o r d e r t o i n d u c ~ a n a d e q u a t e m a t e r n a l

i m m u n e r e s p o n s e , t h e s e p a t i e n t s h a v e b e e n i m m u n i z e d w i t h

p a t e r n a l o r t h i r d p a r t y d o n o r l e u k o c y t e s . 3 , 7 - 1 O

T h e m i x e d l y m p h o c y t e c u l t u r e ( M L C ) . i s a n i n v i t r o

m o d e l f o r a l l e g e n e i c T c e l l p r o l i f e r a t i v e r e s p o n s e t h a t h a s

b e e n u s e d t o d e t e c t b l o c k i n g f a c t o r s i n m a t e r n a l s e r u m .

S e v e r a l i n v e s t i g a t o r s r e p o r t e d a n i n h i b i t o r y s e r u m e f f e c t

u p o n t h e a u t o l o g o u s l y m p h o p r o l i f e r a t i v e r e s p o n s e t o t h e

p a r t n e r 's l y m p h o c y t e s i n w o m e n w i t h s u c c e s s f u l p r e g n a n c y

a n d i t s a b s e n c e i n R S A p a t i e n t s .3, 5 , 6 T h e r e f o r e , i t h a s b e e n

s u g g e s t e d t h a t t h e a b s e n c e o f t h i s f a c t o r m a y p r e d i s p o s e t o

a b o r t i o n .1, 2 , 1~ M o r e o v e r , i t s d e t e r m i n a t i o n h a s b e e n a d o p t e d

a s a p a r a m e t e r í o s e l e c t 3 , 9 , 1 1a n d t o a s s e s s R S A p a t i e n t s t h a t

r e c e i v e l e u k o c y t e t r a n s f u s i o n s . 2 , 9 , 1 2 - 1 4T h i s f a c t o r h a s b e e n

o b s e r v e d i n t r a n s f u s e d R S A p a t i e n t s , 2 , 9 , 1 1 , 1 5b u t ç o n t r o v e r s i a l

r e s u l t s h a v e b e e n r e p o r t e d r e g a r d i n g i t s a s s o c i a t i o n w i t h a

s u b s e q u e n t s u c c e s s f u l p r e g n a n c y . 2 , 1 O , 1 2 , 1 3 , 1 5 - 1 7

T h e M L C i n h i b i t o r y s e r u m f a c t o r h a s b e e n a s s e s s e d

m o s t l y i n n o n - p r e g n a n t w o m e n w i t h a s u c c e s s f u l o b s t e t r i c

DAHEA, S.; FO NSECA, F.; M ATTAA, A.; M USATTI, C.C.; G EABASE DE LIM A, M . -Inhibitory serum facto r of Iym phoproliferative response to.allogeneic cells in pregnancy

R E S U L T S

F i g u r e 1 - P e r c e n t a g e o f p r e g n a n t w o m e n a n d p a t i e n t s w i t h

u n e x p l a i n e d r e c u r r e n t s p o n t a n e o u s a b o r t i o n s ( A S A ) t h a t p r e s e n t e d

m i x e d I y m p h o c y t e c u l t u r e ( M L C ) i n h i b i t o r y s e r u m f a c t o r .

F i s h e r 's e x a c t t e s t : p r i m i g r a v i d a e x 1p r e v i o u s p r e g n a n c y : p

=

0 , 0 4 9 ;

p r i m i g r a v i d a e x p r i m a r y a b o r t e r s : N S ; p r i m i g r a v i d a e x s e c o n d a r y

a b o r t e r s : p=0,01 ;p r i m a r y a b o r t e r s x s e c o n d a r y a b o r t e r s : p=0,01.

A n inhibitory activity on the m ixed lyrnphocyte culture involving the m aternal lyrnphoproliferative response to partner's cells w as observed in the sera of 45% (9/20) ofthe pregnant w om en w ho had at least one parity and in only 8% (1/13) ofthe prim igravidae (p

<

0.05) (F ig. 1). T he frequency of the inhibitory factor in the sera ~ f pregnant w om en w ith one previous pregnancy (3/6) did not differ from that observed in pregnant w om en w ith m ore than one previous pregnancy (6/14). F ifteen patients w ere assessed during the frrst, 14 during the second and four during the third trim ester of pregnancy and the serum factor w as detected w ith the sam e frequency in the three trim esters of pregnancy.

5 0 ' 60

40 30 20 10

%o f s e r a t h a t p r e s e n t e d M L V i n h i b i t o r y f a c t o r o

s e c o n d a r y a b o r t e r s

n=12

3 o rm o r e p r e g n a n c i e s

n=1 4

p r i m a r y a b o r t e r s

n=2 8

autom ated harvester '(T .itertek C ell H arvester) and processed for [3H ]thym idine uptake scintillation counting. T he presence of an inhibitory factor in the serum w as defined as a decrease of at least 50% of the counts in cultures w ith the experim etnal serum ' as com pared to the m ean counts observed w ith the controI serum .

3. Phytohemagglutinin

(PHA) stimulated

cultures:

P eripheraI blood m ononuclear cells (lxl05) w ere incubated in culture rnedium w ith P H A (D ifco, D etroit, U S A ) at a concentration of 10 rng/m I, for three days. T he cultures w ere set up in triplicates, w ith 20% of experim ental or controI sera. O therw ise, the m ethod w as the sam e as the one described for the M L C .

4. Statistical

analysis:

D ata w ere analysed by the F isher's exact test.and a p value < 0,05 w as taken as the leveI of significance.

s e c o n d p r e g n a n c y

n = 6 p r i m i g r a v i d a e

n=1 3

M A T E R I A L A N D M E T H O D S

history and in R S A patients.IO ,1l,13,14,16,18 In the present study w e have assessed the sera of healthy pregnant w om en and of non- transfused R S A patients, in order to determ ine w hether this factor is essential to the developm ent of a norm al pregriancy and to investigate w hether its absence could be considered a relevant param eter for the diagnosis and therapeutic orientation of R S A patients.

1. Patients:

T he study com prised 33 pregnant w om en w ith norm al obstetric histories and 40 non-pregnant w om en w ith recurrent spontaneous abortions (R S A ). T hirteeen of the pregnant w om en w ere prim igravidae and the others had 1 to 6 previous pregnancies. T hey w ere assessed in different trim esters of pregnancy. R S A w as defined as three or .m ore successive m iscarriages in the first trim ester of pregnancy in w om en w ithout evidence of uterine abnorm alities, genital-tract infections, horm onal deficiencies or autoantibodies. C hrom osom e abnorm alities w ere not detected in the patients or their partners. R S A patients w ere classif~ ed as prim ary or secondary aborters on the basis of their obstetric histories. T he w om en w ho had never had a viable pregnancy w ere considered to be prim ary aborters, w hereas w om en w ho had already had a li ve birth or stillbirth w ith m ore than 20 w eeks of gestation w ere classified as secondary aborters.

N one of the w om en assessed in this study had a history of previous blood transfusion. Inform ed consent to participate in the study w as obtained from all couples. T his study w as conducted at the Im m unology L aboratory of tlie A llergy, C linicaI Im m unology and R heum atology D ivision - D epartm ent of P ediatrics of U N IF E S P .

2. Mixed

lymphocyte

culture:

M ononuclear cells w ere separated from peripheraI blood using a F icoll-H ypaque density gradiente T he one-w ay M L C test w as perform ed w ith 1 x 105 _{responder ceIIs and 1 x 10}5

M itom ycin C treated stim ulator cells in R P M I-1640 \S lgm a, S t. L ouis, M o) culture m edium supplem ented w ith H E P E S , L -glutam ine, non-essential am ino acids, sodium bicarbonate and gentam icin. T he cultures w ere carried out in triplicates, in the presence 20% controI serum (pooI of m ale donor sera) or 20% serum being assessed for the presence of an M L C inhibitor factor. A ll sera w ere heat inactivated, at 56° C for 30 m inutes. T he cells w ere incubated at 37° C in a 5% C O₂ atm osphere for 6 days. E ighteen hours before term ination, the cultures w ere labeled w ith 2 m C i of [3H ]thym idine (specific activity 2.48 x

lO "

B q/m m ol). T he cells w ere retrieved using an

1 4 8 7

F ig u re 2 - P e rc e n ta g e o f s e ra th a t p re s e n te d a n in h ib ito ry e ffe c t in a m ix e d Iy m p h o c y te c u ltu re (M L C ) p e rfo rm e d w ith Iy m p h o c y te s fro m H L A c 1 a s s II in c o m p a tib le d o n o rs .

* w ith n o rm a l o b s te tric h is to ry .

fre q u e n c ie s o f s e ra w ith a s u p p re s s iv e e ffe c t w e re o b s e rv e d a m o n g p re g n a n t w o m e n (4 5 % ), s e c o n d a ry a b o rte rs (4 0 % ) a n d n o n -p re g n a n t w o m e n w ith a t le a s t o n e p re v io u s p re g n a n c y (4 3 % ). O n ly 1 o u t o f 9 (1 1 % ) o f th e n u llip a ra e s e ra (p =

0 ,1 1 w h e n c o m p a re d to th e a b o v e m e n tio n e d g ro u p s ) a n d n o n e o f th e m a le s e ra p re s e n te d a s u p p re s s iv e e ffe c t (F ig . 2 ).

% o f s e ra th a t p re s e n te d M L C in h ib ito ry fa c to r

5 0

4 0 30

,2 0

1 0

p re g n a n t w o m e n

n=2 0

o s e c o n d a ry a b o rte rs

n = 5

n o n -p re g n a n t w o m e n w ith p re v io u s p re g n a n c y

n = 7

n u llip a ra e

n = 9

A m o n g R S A p a tie n ts , 2 9 % o fth e p rim a ry a n d 5 8 % o f th e s e c o n d a ry R S A p a tie n ts p re s e n te d th e in h ib ito ry s e ru m fa c to r (F ig . 1 ), w ith a n o v e ra ll in c id e n c e o f 3 8 % . T h e d iffe re n c e in in c id e n c e o f in h ib ito ry fa c to r b e tw e e n p rim a ry a n d s e c o n d a ry a b o rte rs w a s n o t s ta tis tic a lly s ig n ific a n t (p

=

0 .0 9 ). H o w e v e r, th e in c ic i.e n c e o f th e in h ib ito ry fa c to r w a s s ig n ific a n tly h ig h e r a m o n g s e c o n d a ry (p

=

0 .0 1 ), b u t n o t a m o n g p rim a ry a b o rte rs , w h e n c o m p a re d to p rim ig ra v id a e w o m e n . N o s ig n ific a n t d iffe re n c e s w e re d e te c te d re g a rd in g th e in c id e n c e o f th is s e ru m fa c to r in th e p rim a ry o r s e c o n d a ry R S A p a tie n ts w h e n th e y w e re c o m p a re d to p re g n a n t w o m e n w ith a t le a s t o n e p re v io u s p re g n a n c y .

A s u p p re s s iv e e ffe c t o f th e s e ru m o n th e m a te rn a l lu m p h o p ro life ra tiv e re s p o n s e to th ird p a rty ly m p h o c y te s w a s o b s e rv e d in ,4 0 % o f th e n o n -p rim ig ra v id a e p re g n a n t w o m e n , in 2 1 % o f th e p rim a ry a b o rte rs a n d in 2 5 % o f th e s e c o n d a ry a b o rte rs . W ith fe w e x c e p tio n s -, th e s e ra th a t in h ib ite d th e re s p o n s e to th ird p a rty c e lls a ls o in h ib ite d th e re s p o n s e to p a rtn e r's c e lls (T a b le 1 ).

T h e s u p p re s s i v e e ffe c t o f s e ra fro m d iffe re n t s o u rc e s w a s a ls o in v e s tig a te d in a o n e -w a y M L C p e rfo rm e d w ith c e lls fro m tw o H L A (h u m a n le u k o c y te a n tig e n s ) c la s s II in c o m p a tib le d o n o rs , n o t in c lu d in g c e lls fro m th e p a rtn e r o r fro m th e in d iv id u a ls w h o s e s e ra w e re b e in g te s te d . S im ilq r

T a b le 1

P e rc e n ta g e o f p re g n a n t w o m e n w ith a t le a s t o n e p re v io u s p re g n a n c y a n d w o m e n w ith re c u rre n t u n e x p la in e d s p o n ta n e o u s a b o rtio n s (R S A ) th a t

p re s e n te d in h ib ito ry s e ru m fa c to r in m ix e d Iy m p h o c y te c u ltu re s (M L C ) p e rfo rm e d w ith

d iffe re n t c e ll c o m b in a tio n s .

A n in h ib ito ry s e ru m e ffe c t o n th e P H A -in d u c e d ly m p h o p ro life ra tiv e re s p o n s e w a s o b s e rv e d in o n ly

1 0 % (3 /2 9 ) o f th e p re g n a n t w o m e n , a n d in 5 % (1 /2 0 ) o f R S A p a tie n ts .

D IS C U S S IO N

R e sp o n d e r 8 tim u la to r P re g n a n t P rim a ry 8 e co n d a ry ce lls ce lls w o m e n a b o rte rs a b o rte rs

2 0 % ** 8 % ** 2 2 % **

F - fe m a le ; P - p a rtn e r; T - th ird p a rty F m , P m , T m - m y to m ic in tre a te d c e lls

* P e rc e n ta g e o f w o m e n w h o p re s e n te d in h ib ito ry s e ru m ta c to r b o th in c u ltu re s w ith P m a n d w ith T m a s s tim u la to r c e lls

** P e rc e n ta g e o f w o m e n w h o p re s e n te d in h ib ito ry s e ru m ta c to r

b o th in c u ltu re s w ith P a n d w ith T a s re s p o n d e r c e lls .

.F

F

P T

P m T m

F m F m

n = 2 0 n = 1 8 n = 1 2

4 5 % 2 9 % 5 8 %

4 0 % 2 1 % 2 5 0 /0

4 0 % * 1 4 % * 2 5 % *

n = 2 0 n = 1 3 n = 9

2 0 % 1 5 % 2 2 %

3 5 % 8 % 3 3 %

T h e in v e s tig a tio n o f a fa c to r in th e s e ru m o f w o m e n w ith a s u c c e s s fu l p re g n a n c y h is to ry a n d fro m R S A p a tie n ts c a p a b le o f in h ib itin g th e

in vitro

p ro life ra tiv e re s p o n s e o f m a te rn a l to p a te rn a l c e lls (M L C ) h a s b e e n th e fo c u s o f m a n y s tu d ie s p u b lis h e d o v e r th e la s t fe w y e a rs . R e g a rd in g R S A p a tie n ts , th e a b s e n c e o f th is fa c to r w a s c o n s id e re d ,b y M c ln ty re e t a I.3_{a s a c h a ra c te ris tic o f p rim a ry a b o rte rs}

fo r w h o m im m u n iz a tio n w ith th e p a rtn e r's le u k o c y te s w o u ld b e in d ic a te d . O n th e o th e r h a n d , a c c o rd in g to th e s e s a m e a u th o rs , in s e c o n d a ry a b o rte rs , a g ro u p th a t w o u ld b e n e fit fro m h e p a rin th e ra p y , th is fa c to r is o n ly p re s e n t in a fra c tio n o f th e c a s e s . In th e p re s e n t s tu d y a n in h ib ito ry a c tiv ity o f th e s e ru m w a s d e te c te d in 2 9 % o f th e p rim a ry a b o rte rs a n d in 5 8 % o f th e s e c o n d a ry , c le a rly s h o w in g th a t th e a b s e n c e o f th is fa c to r c a n n o t b e . u s e d a s a d is tin c ti v e m a rk e r fo r p rim a ry a b o rte rs .

D A H E R , S .; F O N S E C A , F .; M A T IA R , R .; M U S A T II, C .C .; G E R B A S E D E L IM A , M .

-In h ib ito ry s e ru m fa c to r o f Iy m p h o p ro life ra tiv e re s p o n s e to a llo g e n e ic c e lls in p re g n a n c y

A n o th e r a s p e c t th a t h a s d ra w n a tte n tio n to th e M L C

in h ib ito ry s e ru m fa c to r h a s b e e n its p o te n tia l to p re d ic t

th e o u tc o m e o f a p re g n a n c y fo lIo w in g th e im m u n iz a tio n

w ith p a re n ta lle u k o c y te s . S o m e a u th o rs h a v e fo u n d th a t

th e d e v e lo p m e n t o f th is fa c to r c o u ld b e c o n s id e re d

p re d ic tiv e o f a s u c c e s s fu l p re g n a n c y 2 ,4 ,5 ,8 ,1 1 ,1 2 , w h ile o th e rs

h a v e n o t fo u n d s u c h a c o rre la tio n . 1 0 ,1 4 -1 6 W e re a s o n e d th a t

if th is fa c to r is b io lo g ic a lIy im p o rta n t fo r th e d e v e lo p m e n t

o f a s u c c e s s fu l p re g n a n c y , it s h o u ld re g u la rly b e d e te c te d

in th e s e ru m fro m p re g n a n t w o m e n . T h e p re s e n t d a ta s h o w

th a t th is is n o t th e c a s e , s in c e th e in h ib ito ry fa c to r w a s

d e te c te d in o n ly 1 o u t o f 1 3 p rim ig ra v id a e , a lI o f w h o m

h a v e a lre a d y d e liv e re d a h e a lth y b a b y . F u rth e rm o re , a

s im ila r in c id e n c e o f th e in h ib ito ry fa c to r w a s o b s e rv e d in

p re g n a n t w o m e n w h o h a d a t le a s t o n e p re v io u s p a rity

(4 5 % ) a n d in s e c o n d a ry a b o rte rs (5 8 % ). W e b e lie v e th a t

th e s e re s u lts , ta k e n to g e th e r, s u p p o rt th e c o n te n tio n th a t

th e in h ib ito ry fa c to r s h o u ld b e re g a rd e d a s a n

e p ip h e n o m e n o n o f p re g n a n c y ra th e r th a n a n im p o rta n t

fa c to r fo r fe ta l s u rv iv a l, a s h a s a lre a d y b e e n s u g g e s te d .5 ,18

T h e n a tu re o f th e M L C in h ib ito ry fa c to r is n o t y e t

fu lly c la rifie d . It h a s b e e n s u g g e s te d th a t th e b lo c k in g

a c tiv ity m a y b e re la te d to a n tib o d ie s d ire c te d a g a in s t H L C

a n tig e n s o r a g a in s t tro p h o b la s t a n tig e n s th a t c ro s s -re a c t

w ith H L A a n tig e n s , o r c o u ld re p re s e n t a n ti-id io ty p e

a n tib o d ie s d ire c te d to th e T c e lI re c e p to rs (T C R ) in v o lv e d

in a lIo re c o g n itio n . O u r s tu d y w a s n o t s p e c ific a lly d e s ig n e d

to s tu d y th e n a tu re o f th e in h ib ito ry fa c to r, b u t w e h a v e

a d d re s s e d th e q u e s tio n c o n c e rn in g its s p e c ific ity in

b lo c k in g th e re s p o n s e o f th e p re g n a n t o r R S A w o m e n

a g a in s t th e ir p a rtn e rs ' c e lIs . R e s u lts h a v e s h o w n th a t th e

m a jo rity o f th e s e ra th a t in h ib ite d th e c u ltu re s w ith th e

p a rtn e r c e lls a ls o s h o w e d in h ib itio n in c u ltu re s w ith o th e r

c e ll c o m b in a tio n s (T a b le 1 a n d F ig . 2 ), c o n firm in g

o b s e rv a tio n s o f o th e r a u th o rs .2,6 It is in te re s tin g to n o te

th a t n o in h ib ito ry a c tiv ity w a s d e te c te d in a n y o f th e m a l e

s e ra te s te d , a s a lre a d y re p o rte d in m e d ic a llite ra tu re .6

W h e n

th e w o m e n 's s e ra w e re a s s a y e d in P H A -s tim u la te d c e ll

c u ltu re s , o n ly a fe w (th re e o u t o f 2 9 p re g n a n t a n d o n e o u t

o f 2 0 R S A ) s h o w e d a n in h ib ito ry e ffe c t. T h u s it a p p e a rs

th a t th e in h ib ito ry fa c to r s o m e h o w in te rfe re s w ith

a llo re c o g n itio n o r, a lte rn a tiv e ly , th a t is is n o t p o w e rfu l

e n o u g h to b lo c k th e re s p o n s e to m ito g e n s th a t re p re s e n t a

s tro n g e r s tim u lu s th a n a llo g e n e ic c e lls .

C O N C L U S I O N

O u r d a ta s u g g e s t th a t th e M L C in h ib itin g fa c to r s h o u ld

b e re g a rd e d a s a c o n s e q u e n c e o f p re g n a n c y ra th e r th a n a n

e s s e n tia l fa c to r fo r fe ta l s u rv i v a I. T h e re fo re , w e b e lie v e th a t

th is te s t, w h ic h is c o s tly , is n o t in fo rm a tiv e e ith e r re g a rd in g

th e d e c is io n to im m u n iz e R S A w o m e n w ith p a re n ta l

le u k o c y te s o r a s a p re d ic to r o f a s u c c e s s fu l p re g n a n c y in

w o m e n s u b m itte d to im m u n iz a tio n . It is th e re fo re o u r

o p in io n th a t th e o n ly im m u n o lo g ic a l te s t th a t s h o u ld b e

p e rfo rm e d in R S A p a tie n ts th a t a re b e in g c o n s id e re d fo r

e n te rin g a n im m u n iz a tio n p ro to c o l is th e s e a rc h fo r

ly m p h o c y tq to x ic a n tib o d ie s a g a in s t th e c e lls (c ro s s m a tc h )

th a t w ill b e u s e d fo r im m u n iz a tio n . A p o s itiv e c ro s s m a tc h

s h o u ld c o n tra in d ic a te th e im m u n iz a tio n in o rd e r to a v o id

h y p e rs e n s itiz a tio n a n d a ls o b e c a u s e , a s n o te d b y s o m e

a u th o rs , 1 0a p o s itiv e c ro s s m a tc h is a lre a d y a n in d ic a tio n th a t

a n im m u n e re s p o n s e to p a te rn a l a n tig e n s h a s o c c u rre d .

A C K N O W L E D G E M E N T S

T h e a u th o rs th a n k D r. L e ila F a ra h fo r p e rfo rm in g

th e c h ro m o s o m e a n a ly s is .

S ã o P a u l o M e d i c a i J o u r n a l / R P M 1 1 5 ( 4 ) : 1 4 8 5 - 1 4 8 9 , 1 9 9 7 D A H E R , S . ; F O N S E C A , F . ; M A T I A R , R . ; M U S A T T I , C . C . ; G E R B A S E D E L I M A , M .

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2. B eer A E , Sem prini A E , Z hu X , Q uebbem an JF. Pregnancy outcom e in hum an couples w ith recurrent spontaneous abortions: H L A antigen profiles; H L A antigen sharing; fem ale serum M L R blocking factors, and paternalleukocyte im m unization. E xp C lin Im m unogenet 1985; 2: 137-153. 3. M clntyre JA , Faulk W P L aboratory and clinicaI aspects of

research in chronic spontaneous abortion. D iag Im m unol 1985; 3: 163-170.

4. B eer A E , Q uebbem an JF, H am azaki Y , Sem prini A E . Im m unotherapy of recurrent spontaneous abortion. In: G ill T J, W egm ann T G , eds. Im m unoregulation and fetal survivaI. N ew Y ork : O xford U niversity Press. 1987:286 - 299. 5. T haler C J, M clntyre J A . Fetal w astage and nonrecognition

in hum an pregnancy. Im m unol A llergy 1990; 10:79-101. 6. ScottJR , R oteN S, B ranch D W . Im m unologic aspects of recurrent

abortion and fetal death. O bstet G ynecol1987; 70: 645-656. 7. H ill JA . Im m unologic m echanism s of pregnancy m aintenance and failure: a critique of theories and therapy. A m J R eprod Im m unol 1990: 22: 33-42

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11. U nander A M , L indholm A . T ransfusion of leukocyte-rich erythrocyte concentrates: A successful treatm ent in selected cases ofhabitual abortion. A m J O bstet G ynecol1986; 154: 516-520. 12. T akakuw a K ., G oto S., H asegaw a I. et aI. R esult of

im m unotherapy on patients w ith unexplained recurrent abortion: A beneficial treatm ent for patients w ith negative blocking antibodies. A m J R eprod Im m unol1990; 23: 37- 41. 13. Sugi T , M akino T , M aruyam a T , K im K lizuka R . A

possible m echanism of im m unotherapy for patients w ith recurrent spont'lneous abortions. A m J R eprod Im m unol 1991; 25: 185-189.

14. Illeni M T , M arelli G , Parazzini F, A caia B , B occiolone L , B ontem pelli M et aI. Im m unotherapy and recurrent abortion:

a random ized clinicaI triaI. H um R eprod 1994; 9: 1247-1249. 15. H o H G , G ill T J, H sieh H J, Jiang JJ, L ee T Y , H sieh C Y .

Im m unotherapy for recurrent spontaneous abortion in a chinese population. A m J R eprod Im m unol1991; 25:10-15. 16. C ow chock FS, Sm ith JB , D avid S, Scher J, B atzer F, C orson S. - Paternal m ononuclear cell im m unization therapy for repeated m iscarriage: predictive variables for pregnancy success. A m J R eprod Im m unol 1990; 22: 12 -17. 17. C lark D and D aya S. T rials and tribulation in the treatm ent

of recurrent spontaneous abortion. A m J R eprod Im m unol 1991; 25: 18-24.

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D A H E R , S .; F O N S E C A , F .; M A lT A R , R .; M U S A T T I, C .C .; G E R B A S E D E L IM A , M .

-In h ib ito r y s e r u m fa c to r o f Iy m p h o p r o life r a tiv e r e s p o n s e to a llo g e n e ic c e lls in p r e g n a n c y